• Title/Summary/Keyword: Cell suspension culture

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Selection of Acid Tolerant Red Clover Cell Line on the Cellular Level II. Effect of some factors affecting suspension culture and acid tolerant cell selection (내산성 레드 클로-버의 배양세포수준에서의 선발 II. 레드 클로-버의 현탁배양 및 내산성 세포의 선발)

  • Son, Dae-Young-Son;Lee, Soon-Hee-Lee;Jo, Jin-Gi
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.13 no.1
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    • pp.1-6
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    • 1993
  • This experiment was conducted to select acid tolerant cells from red clover suspension culture and obtained following results. The most effective condition of plant growth regulators for suspension culture of red clover cells was 2 mg/l 2,4-D as auxin source plus 0.5 mg/l BAP as cytokinin source. Among serveral basal media, PC medium brought the best result. The most suitable EMS concentration for the mutated cells tolerant to acid was 1 % with fours treatment and average 27 colonies/plate were selected. The frequency of the occurence of mutants tolerant to acid in 1 % EMS with four hours treatment was 8.9 $\times$ 10$^{-5}$ which was 100 times larger than that of control. The total colonies selected initially were 176 but the survived colonies after two subcultures with 4 weeks interval in a selection medium were 44.

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Botryococcus braunii 배양에서 탄화수소의 two-stage 동시추출공정

  • An, Jin-Yeong;Choe, Jeong-Gyu;Sim, Sang-Jun;Kim, Byeong-U
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.229-232
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    • 2000
  • The carbon dioxide, nitrogen, and phosphate removals from wastewater using microalgae have extensively been studied. Especially, the green colonial algae Botryococcus braunii is characterized by unusual high hydrocarbon contents, ranging from 15 to 75% of dry weight, as long-chain unsaturated hydrocarbons. These hydrocarbons suggest that the possibility of renewable biofuels to be converted into useful fuels such as gasoline by simple catalytic cracking. The poor recovery (18 - 32%) of hydrocarbon from B. braunii culture in two-phase bubble column seems to be caused by insufficient mixing between two phases, which was operated using only aeration on the narrow interface between hydrophobic solvent and cell suspension. In addition, hydrocarbon was entrapped tightly in cell-matrix (formed by exopolysaccharide) of algal colony, which make difficult to extract using two-phase system. In order to overcome low recovery efficiency, two-stage extraction culture system including culture vessel and two-phase separator is now under development, resulted improving contact between solvent phase and cell suspension. Hydrocarbon recovery using this process was more than two times as that using two-phase extraction culture.

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Derivation of Neural Precursor Cells from Human Embryonic Stem Cells

  • Kim Sehee;Hong Ji Young;Joo So Yeon;Kim Jae Hwan;Moon Shin Yong;Yoon Hyun Soo;Kim Doo Han;Chung Hyung Min;Choi Seong-Jun
    • Reproductive and Developmental Biology
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    • v.28 no.4
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    • pp.247-252
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    • 2004
  • Human embryonic stem (ES) cells are derived from the inner cell mass of the preimplantation embryo. Human ES cells have the capacity to differentiate into various types of cells in the body. Human ES cells are indefinite source of cells for cell therapy in various degenerative disorders including neuronal disorders. Directed differentiation of human ES cells is a prerequisite for their clinical application. The objective of this study is to develop the culture condition for the derivation of neural precursor cells from human ES cells. Neural precursor cells were derived from human ES cells in a stepwise culture condition. Neural precursor cells in the form of neural rosette structures developed into neurospheres when cultured in suspension. Suspension culture of neurospheres has been maintained over 4 months. Expressions of nestin, soxl, sox2, pax3 and pax6 transcripts were upregulated during differentiation into neural precursor cells by RT-PCR analysis. In contrast, expression of oct4 was dramatically downregulated in neural precursor cells. Immunocytochemical analyses of neural precursor cells demonstrated expression of nestin and SOX1. When induced to differentiate on an adhesive substrate, neuro-spheres were able to differentiate into three lineages of neural systems, including neurons, astrocytes and oligo-dendrocytes. Transcripts of sox1 and pax6 were downregulated during differentiation of neural precursor cells into neurons. In contrast, expression of map2ab was elevated in the differentiated cells, relative to those in neural precursor cells. Neurons derived from neural precursor cells expressed NCAM, Tuj1, MAP2ab, NeuN and NF200 in immunocytochemical analyses. Presence of astrocytes was confirmed by expression of GFAP immuno-cytochemically. Oligodendrocytes were also observed by positive immuno-reactivities against oligodendrocyte marker O1. Results of this study demonstrate that a stepwise culture condition is developed for the derivation of neural precursor cells from human ES cells.

Plant Regeneration from Cryopreserved Embryogenic Cell Suspension Cultures of Cucumber (초저온 보존된 오이 배발생세포 현탁배양으로부터 식물체 재분화)

  • Kim, Seok-Won;In, Dong-Soo;Jung, Won-Joong;Woo, Je-Wook;Jung, Min;Yoo, Jang-Ryul
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.6
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    • pp.501-505
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    • 1998
  • Conditions for high frequency plant regeneration from cryopreserved embryogenic cell suspension cultures derived from hypocotyl explants of cucumber (Cucumis sativus L.) are described. Cells cryoprotected with a mixture of 2 M DMSO and 0.4 M sucrose exhibited a regeneration frequency of 85%. However, cells cryoprotected with different concentrations of glycerol showed no regeneration after cryopreservation. Pretreatment of cells in a high osmotic medium was not necessary to the process. Upon transfer to MS medium supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid, regenerated calli gave rise to numerous somatic embryos, then underwent development into plantlets.

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Effects of Inoculum Density and Basal Media on Cell Growth and Taxol Production in Taxus Cell Suspension Cultures (주목 세포배양에서 초기 접종농도와 기본배지가 세포증식과 Taxol 생산에 미치는 영향)

  • 황용순;김석우
    • KSBB Journal
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    • v.11 no.5
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    • pp.600-605
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    • 1996
  • Optimum inoculum concentration for the production of taxol was determined in Taxus brevifolia and Taxus cuspidata cell suspension cultures. By fresh weight, 2.5, 5, 7.5, 10 g/flask of cells were inoculated and cell growth as well as taxol production were examined. In both Taxus cell cultures, the higher the inoculum concentration, the shorter the length of the lag period. The optimum inoculum concentration for taxol production was found to be 5 g/flask. To produce taxol in large quantity, utilization of proper medium was thought to be important. In case of using a production medium with 6% sucrose, taxol production was noticed. Its level reached the maximum at the 9th day of culture and decreased afterwards. However, taxol was not detected from cell cultures in growth medium.

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Biosynthesis and Metabolism of Vitamin C in Suspension Cultures of Scutellaria baicalensis

  • Ahn, Young-Ock;Kwon, Suk-Yoon;Lee, Haeng-Soon;Park, Il-Hyun;Kwak, Sang-Soo
    • BMB Reports
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    • v.32 no.5
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    • pp.451-455
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    • 1999
  • The concentrations of L-ascorbic acid (AsA, ascorbate, vitamin C) and its biosynthetic and metabolically-related enzymes such as L-galactono-1,4-lactone dehydrogenase (GLDase), ascorbate peroxidase (APX), and ascorbate oxidase (ASO) were investigated in suspension cultures of Scutellaria baicalensis. Cells growing from 4 days after subculture (DAS) to 9 DAS and from 16 DAS to 19 DAS showed a diauxic growth, and then growth rapidly decreased with further culturing. The AsA content slowly increased to 19 DAS, reached a maximum at 21 DAS (ca $120\;{\mu}g/g$ dry cell wt), and then rapidly decreased with further culturing. GLDase and ASO activity were well correlated with the cell growth curve, showing a maximum at 19 DAS, whereas APX activity showed a good correlation with the changes in AsA content, showing a maximum at 21 DAS. The total ascorbate contents (reduced form, AsA, and oxidized form, dehydroascorbate, DHA) were markedly enhanced at 10 DAS when L-galactose and L-galactono-1,4-lactone (25 mM) were added to SH medium supplemented with 20 g/l sucrose at 9 DAS, by 5.5 and 6.8 times, respectively. DHA composed more than 90% of the total ascorbate contents in suspension cultures of S. baicalensis, even though the ratio of reduced to oxidized form slightly varied with cell growth stage. The results indicate that L-galactose and L-galactono-1,4-lactone are effective precursors of AsA in cell cultures of S. baicalensis, and that in vitro cultured cells provide suitable biomaterials for the study of biosynthesis and metabolism of AsA.

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